LEGUME INDIAN BEAN (PHASEOLUS VULGARIS) PARTIALLY PURIFIED ANTI-NUTRITIONAL PROTEIN FACTOR EXHIBIT ANTI-OXIDANT, IMMUNOMODULATORY AND ANTI-CANCER PROPERTIES

HEMAVATHI AVALAPPA; SHRUTHISHREE D. PADIYAPPA; NIRANJANA PATHAPPA; PRABHAKAR T. BETTADATUNGA; SIDDANAKOPPALU N. PRAMOD

doi:10.22159/ijpps.2023v15i2.46831

Authors

HEMAVATHI AVALAPPA Department of Biochemistry, Kuvempu University, Shankaragatta 577451, Shivamogga, Karnataka, India
SHRUTHISHREE D. PADIYAPPA Department of Biochemistry, Kuvempu University, Shankaragatta 577451, Shivamogga, Karnataka, India
NIRANJANA PATHAPPA Department of Biochemistry, Kuvempu University, Shankaragatta 577451, Shivamogga, Karnataka, India
PRABHAKAR T. BETTADATUNGA Molecular Biomedicine Laboratory, Postgraduate Department of Biotechnology, Sahyadri Science College, Kuvempu University, Shivamogga 577203, Karnataka, India
SIDDANAKOPPALU N. PRAMOD Food Allergy and Immunology Laboratory, Department of Studies in Food Technology, Davangere University, Shivagangotri, Davangere 577007, Karnataka, India

DOI:

https://doi.org/10.22159/ijpps.2023v15i2.46831

Keywords:

Anti-nutritional factor, Antioxidant, Anti-angiogenic, Anti-cancer, Immunomodulatory

Abstract

Objective: This study was aimed to access one of the hidden ANF in Indian bean and assessment of its antioxidant, immunomodulatory, antiangiogenic, and anti-cancer properties.

Methods: Indian beans were processed and evaluated for the presence of ANF through HA activity. Bean-ANF was partially purified by ammonium sulfate precipitation (ASP) and dialysis. Obtained Partially Purified bean-ANF (PPb-ANF) resolved on 12% SDS-PAGE, analyzed protease digestibility. The immunogenicity of PPb-ANF was characterized by producing polyclonal antibodies and checked for the presence of natural antibodies in healthy human sera. Pharmacological studies of PPb-ANF were assessed through anti-oxidant, immunogenic, anti-angiogenic and anti-cancer assays.

Results: Partially purified Bean-ANF (PPb-ANF) showed significant specific HA activity up to 150 Units/mg compared to crude extract 9.3 of Units/mg. The PPb-ANF was identified to have more than 90% homogeneity in purity with an observed M W of 30-32 kDa. PPb-ANF showed significant pepsin digestive stability up to 2 h at 37 ᵒC and appears digestive resistant. Rabbit produced anti-PPb-ANF IgG polyclonal antibody on immunization and exhibited immunogenic properties. Natural human IgG and IgE antibodies were found in the non-atopic human sera against PPb-ANF, indicative of its immunogenicity. Pharmacological studies of PPb-ANF confirmed its potent antioxidant activity. At 400 µg of PPb-ANF showed the highest around 90% DPPH activity and at 150 µg of PPb-ANF showed 89% inhibitory lipid peroxidation activity. The PPb-ANF showed 70% (3 μg) and 40% (1.5 μg) inhibitory in blood vessel density compared to control and exhibited significant anti-angiogenic activity. The PPb-ANF showed a significant (*p<0.05.) reduction in solid tumor weight upon dose-2 treatment (50 mg/kg body weight) and also increased lifespan up to 70 d compared with the control tumor model.

Conclusion: PPb-ANF was identified as digestive-resistant, hemagglutinin with a molecular weight of 30-32 kDa. The protein is immunogenic and has potential antioxidant, antiangiogenic and anti-cancer properties. Further purification and mechanistic-based pharmacological characterization might explore this protein as a potential anti-cancer molecule.

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